Method and system for producing high purity vancomycin hydrochloride

ABSTRACT

A method is provided for preparing spray dried powder containing vancomycin hydrochloride. The method comprises providing a vancomycin hydrochloride solution with a chromatographic purity of at least 95%, adding an excipient to the vancomycin hydrochloride solution to form a mixture solution of the vancomycin hydrochloride solution and the excipient, concentrating the mixed solution of the vancomycin hydrochloride solution and the excipient to form a 20% to 30% vancomycin concentrate, filtering the vancomycin concentrate to form a final filtrate, and spray drying the final filtrate to form a spray dried vancomycin hydrochloride powder with EP impurity B level of not more than 1.5%.

CROSS-REFERENCE TO RELATED APPLICATION

This application is based on and claims priority under 35 U.S.C. 119from Chinese Patent Application No. 201310085761.7, filed Mar. 15, 2013,titled “Spray Dry Powder and Industrial Preparation of VancomycinHydrochloride,” which is hereby incorporated herein by reference in itsentirety.

FIELD OF THE DISCLOSURE

The present disclosure relates to a system and a method for producinghigh purity vancomycin hydrochloride.

BACKGROUND OF THE DISCLOSURE

Vancomycin hydrochloride is an amphoteric glycopeptide antibioticproduced by fermentation of the microorganism Nocardia orientalis (orAmycolatopsis orientalis) under controlled conditions, which has amolecular formula C₆₆H₇₅Cl₂N₉O₂₄.HCl and a molecular weight of 1.486.Commercial distribution of vancomycin hydrochloride began at the end of1950s and its chloride form has been used clinically ever since.Lyophilized vancomycin hydrochloride is a white or almost white powder.It is hygroscopic, freely soluble in water, and slightly soluble inalcohol. When mixed with water (5%, w/w), vancomycin hydrochloride has apH between 2.5 and 4.5.

Vancomycin hydrochloride acts by binding the C-terminal D-Ala-D-Alapeptides, which inhibits the synthesis of cell walls and also changesthe permeability of cell membranes as well as synthesis of RNA.Vancomycin hydrochloride is particularly used for the initial treatmentof serious or severe infections caused by staphylococci resistant toβ-lactam antibiotics as well as in patients who are penicillin-sensitiveor do not respond to penicillin or cephalosporine. Vancomycinhydrochloride is commercially available in oral (solution andcapsules/pulvules) and parenteral (sterile intravenous solution invials) forms.

Vancomycin hydrochloride, alone or in combination with otheraminoglycosides, is useful in treating staphylococcal, streptococcal,enterococcal or diphtherial endocarditis. An indication of oralvancomycin therapy includes the treatment of pseudomembranous colitiscaused by staphylococci when it is unresponsive to vancomycin forinjection. Vancomycin for injection may be applicable to all of theother indications.

A vancomycin molecule is composed of two basic structures, including asaccharide group, α-o-vancosamine-β-o-glucosyl, and a heptapeptidebackbone. The structure of vancomycin determines its instability and thecase with which it may be degraded under acidic conditions, alkalineconditions and/or high temperature conditions. Normally, degradationproducts have no biological activity, so side effects may be reducedwhen impurity levels are significantly reduced.

Desvancosaminyl vancomycin and aglucovancomycin are degradation productsthat result from the loss of the disaccharide moiety and the vancosaminesugar, respectively, under acidic and high temperature conditions.Vancomycin can be degraded into another degradation product,desamidovancomycin, by hydrolytic loss of ammonia in weak acidconditions. Desamidovancomycin exists in two isomeric forms as shown inFIGS. 1 and 2. The forming mechanism of desamidovancomycin has beendescribed in publications, such as, for example, “Vancomycin degradationproducts as potential chiral selectors in enantiomeric separation ofracemic compounds,” by Alireza Ghassempour, Journal of Chromatography A,1191 (2008) 182-187. In this publication, the authors deduced that thereare two pathways for vancomycin to succinimide and its conversion todesamidovancomycin, as described in FIG. 3.

Industrial methods for preparing vancomycin hydrochloride have beenknown for some time. For instance, U.S. Pat. No. 3,067,099 discloses amethod of producing vancomycin through cultivation of avancomycin-producing strain of Streptomyces Orientalis under aerobicconditions in a culture medium containing assimilable sources ofcarbohydrate, organic nitrogen and inorganic salts. Separation andpurification processes for separating vancomycin hydrochloride fromfermentation broth have also been disclosed in literature and patentdocuments. U.S. Pat. No. 4,440,753 describes an example of an isolationmethod and purification process for forming precipitate by isopropanol,ethanol or acetone. U.S. Pat. No. 4,868,285 discloses an example of anisolation method and filtration process to collect a compound ofimidazole and vancomycin. However, in many of these processes, formednantokite or imidazole compound may decompose and contaminate the finalvancomycin product. Further, slurry may form when using isopropanol,ethanol or acetone to isolate vancomycin, which is hard to filter.

U.S. Pat. No. 5,853,720 discloses a process for purifying vancomycinhydrochloride that combines the preparative chromatography on a silicagel column containing an alkaline water-methanol mobile phase and theprecipitation with ethanol from a salt-water-ethanolic solution. Theprocess produces vancomycin hydrochloride solid by lyophilization, spraydrying or precipitation of pH 3, 100 g/L concentrate, which is obtainedthrough a series of operations like micro-filtration of vancomycinbroth, then macro porous resin adsorption and elution, concentration anddesalting, discoloring with activated carbon. The vancomycin solution isspray dried at an inlet temperature of 115-130° C. and an outlettemperature of 85±5° C. Usually, the content of water in vancomycinhydrochloride is about 4%, therefore it has been practice toadditionally dry vancomycin hydrochloride in a rotation vacuum dryer ata temperature of 45-50° C. to obtain a dry solid product. This patentmentions that a chromatographic purity of vancomycin hydrochloridesolution is about 93%. However, the patent does not appear to indicatethe chromatographic purity and impurity levels of the final dry product.Experts and experienced technicians can appreciate that chromatographicpurity of the obtained vancomycin hydrochloride product may decreasesignificantly after spray drying under high temperature conditions andvacuum drying at medium temperature conditions, resulting in increasedimpurity level and darker product color.

U.S. Pat. No. 7,018,804 discloses a method for preparing high purityvancomycin hydrochloride. In this patent, vancomycin hydrochlorideconcentrate with a HPLC purity of not less than 95% based on the EPmethod is obtained through a series of column purification processes,including strong acid type ion exchange resin chromatography, weak basetype ion exchange resin column and aluminum oxide column chromatography,and hydrophobic resin chromatography. The final vancomycin hydrochlorideproduct is obtained through precipitation by adding multiple (e.g., 5)times volumes of acetone and drying the resultant solution under vacuumconditions at a temperature of 40° C. It is note that this drying methodwould likely elevate the impurity level in the final product. Moreover,using the described method, residual solvents can't be removedcompletely to meet the corresponding requirements of, e.g., theInternational Conference on Harmonisation of Technical Requirements forRegistration of Pharmaceuticals for Human Use (ICH).

M. Nieto and H. R. Perkins, in their publication, “PhysicochemicalProperties of Vancomycin and Iodovancomycin and their Complexes withDiacetyl-L-lysyl-D-alanyl-D-alanine,” Biochem. J. (1971) 123, 773-787,observed aggregation of vancomycin in an aqueous solution. The structureof aggregated glycopeptides remained uncharacterized. The publicationdescribed that aggregation is influenced by many factors such asionization degree of phenol, hydrogen bond, etc.

Through addition of appropriate excipients such as surfactants orcarbohydrates to reduce the forming of aggregates, the limitingconcentration of vancomycin hydrochloride in water can be increased.

Due to thermal instability of vancomycin hydrochloride, the solution istypically prepared at a relative low temperature to avoid an increase inimpurity level. Even if the solution is dried at a low temperature andunder high vacuum conditions, the chromatographic purity of vancomycinhydrochloride will inevitably decrease. The lyophilization at lowtemperature will cause chromatographic purity to decrease by about 0.5%or more. For instance, the purity level of the solution will decrease bymore than 1.0% through vacuum drying at 40° C. within 8 hours. Themainly-increased impurity is desamidovancomycin.

U.S. Pat. No. 6,001,800 discloses a method for preparing spray driederythropoietin, and the dry erythropoietin powder produced thereby. Thepatent discloses that through addition of mannitol, glycine and sodiumcitrate as stabilizers and dispersants, active substances withappropriate ranges of particle size can be obtained.

U.S. Pat. No. 6,479,049 discloses a method and composition for the drypowder formulation of cytokines, especially interferons. The patentmentions interferon, to which carbohydrates, polypeptides and aminoacids are added as a carrier. Appropriate excipients are added toimprove product stability.

Excipients were reported to be used in vancomycin hydrochlorideintravenous infusion in order to improve vancomycin stability andsolubility such as Meek, a lyophilized generic product containing 100 mgeach of D-mannitol and PEG 400 per 500 mg of vancomycin hydrochlorideinjection. Moreover, it was also reported that Meek was less nephrotoxicthan a conventional preparation without addition of excipients, as notedby Naoko Hodoshima, Drug Metab. Pharmacokin. 19(1): 68-75 (2004).

U.S. Patent Application Publication No. US 2013/0009330 discloses apreparative method of a stable and easily-soluble powdered vancomycinformulation for injection through a spray drying process afterdissolving a mixture of 10-20% by weight vancomycin HCl, about 2-4% byweight PEG, and 2-4% by weight mannitol. However, impurity B specifiedin European Pharmacopeia can't be significantly controlled by using thisformulation. Moreover, the concentration of the vancomycin hydrochloridesolution is limited at 10-20%, and it is not easy for further filling ofspray dried powder due to low density.

Presently, drying methods used for drying vancomycin hydrochlorideinclude lyophilization, vacuum drying and spray drying.

Lyophilization tends to be the main drying process for the production ofboth preparations and active substances. During lyophilization of activesubstances, it is difficult to ensure application of aseptic technologyat each stage of the production process, including, e.g., duringloading, unloading and powder collection processes, thereby making itvery difficult to obtain sterile active substances. Therefore, it isfrequently necessary to repeatedly dissolve non-sterile activesubstances and lyophilize the substances to get sterile powder forinjection in vials. This inefficient production process results in largeinvestments in specialized equipment, low production yields, poorefficiency and high operation costs, which may not be applicable for theproduction of large strengths above, e.g., 10 g/vial because of the highratio of vials that tend to break during the process, as well as theundesirably long lyophilization cycle. The prolonged production cycletends to affect product quality due to a higher impurity level.

If using vacuum drying, as a heat-sensitive substance, vancomycin shouldbe dried at a low temperature and under high vacuum conditions. However,because vancomycin tends to combine with water and some polar solvent,it is hard to completely remove the water or the polar solvent. Asignificant problem in vacuum drying is that the residual solvent levelcan't meet the ICH's requirements. Moreover, prolonged drying time willresult in a higher impurity level.

There has been discussion of using spray drying in industrial productionof vancomycin hydrochloride under high temperature conditions. In spraydrying, a vancomycin hydrochloride fluid solution is transformed into adry-form product by spraying the solution into, e.g., a heat dryingdevice. The resultant spray-dried product is typically in powder form.Spray drying, however, can result in degradation of vancomycinhydrochloride. During preparation of the vancomycin chloride solution,vancomycin chloride concentration levels can exceed 20%, therebyincreasing the viscosity of the solution and forming an aggregate (e.g.,semi-solid jelly) or precipitate that may block equipment, e.g.,production filters. The implementation of 15% vancomycin hydrochlorideconcentration levels has been reported, such as, e.g., in US2013009330.

Solution stability should be considered since the concentrate may bestored for more than twenty-four hours because additional preparationtime may be required for each operation. There exists the unsolvedproblem in industrial production on how to ensure stability ofvancomycin hydrochloride solution without increasing impurity andforming precipitate within, for example, forty-eight hours. Since thetemperature during spray drying may exceed above 80° C., ensuring thestability of vancomycin hydrochloride may be a barrier to production.

A further problem with spray drying is that the spray dried vancomycinpowder will be filled directly into, e.g., vials. Accordingly, theprocess should allow for reconstitution time at least as fast as thelyophilized formulations. Spray dried vancomycin tends to have smallerparticle sizes and the reconstitution time tends to be longer than thatof the loose-structured lyophilized product.

In order to reduce the possibility of side effects, a highchromatographic purity of antibiotics is very important in certainapplications, which cannot be achieved by hitherto purificationprocesses.

The present disclosure provides a novel system and a novel method forproducing high purity and high potency vancomycin hydrochloride thatsolve the afore-discussed problems.

SUMMARY OF THE DISCLOSURE

According to one non-limiting example of the disclosure, a system and amethod are provided for producing high purity and high potencyvancomycin hydrochloride from a non-sterile or sterile vancomycinhydrochloride substance with a low impurity level. According to thepresent disclosure, a non-sterile or sterile vancomycin hydrochloridedrug substance with low impurity level can be obtained through afermentation process of Amycolatopsis Oriertalis on appropriate mediacontaining carbon, nitrogen and inorganic salts at appropriatecultivation conditions to obtain vancomycin broth, followed by a seriesof purification steps to obtain above 20% (w/w) vancomycin hydrochlorideultra-concentrate, as well as final evaporation at high temperature in ashort time to remove solvents. This obtained sterile vancomycinhydrochloride drug substance can be directly subdivided to powder forinjection in patients.

The present disclosure provides a method to prepare spray driedvancomycin hydrochloride powder. This method includes using vancomycinhydrochloride solution with a chromatographic purity of not less than95%, adding stabilizer and solubilizer to improve solution stability, aswell as to increase its concentration to 20-30%, and spray drying thesolution to form spray dried vancomycin hydrochloride powder with aimpurity B (specified in EP monograph) level of not more than 1.5%.

The disclosure provides many advantageous over existing systems andprocesses for making vancomycin hydrochloride. For instance, thedisclosure resolves, among other things, the solution stability ofvancomycin hydrochloride at high concentrations by increasing theconcentration to 20-30%, which significantly improves efficiency of thespray drying process. The higher concentration levels also improvequality, dissolving characteristics, bulk density of the obtained drugsubstance powder, impurity level control, yield to realize large scaleproduction, while reducing time and cost requirements of productionoperations.

According to an aspect of the disclosure, a method is provided forpreparing spray dried powder containing vancomycin hydrochloride. Themethod comprises: providing a vancomycin hydrochloride solution with achromatographic purity of at least 95%; adding an excipient to thevancomycin hydrochloride solution to form a mixture solution of thevancomycin hydrochloride solution and the excipient; concentrating themixed solution of the vancomycin hydrochloride solution and theexcipient to form a 20% to 30% vancomycin concentrate; filtering thevancomycin concentrate to form a final filtrate; and spray drying thefinal filtrate to form a spray dried vancomycin hydrochloride powderwith EP impurity B level of not more than 1.5%.

The method may further comprise transferring the spray dried vancomycinhydrochloride powder with EP impurity B level of not more than 1.5% intoa gamma irradiated single use bag.

The excipient may comprise at least one of: a stabilizer; a solubilizer;a saccharide; a polyol; polyethylene glycol-400 (PEG-400); and asurfactant.

The method of providing the vancomycin hydrochloride solution with thechromatographic purity of at least 95% may comprise dissolving avancomycin hydrochloride active pharmaceutical ingredient (API) inpurified water.

The mixture solution of the vancomycin hydrochloride solution and theexcipient may comprise: about 5-35% by weight of saccharide; about10-50% by weight of polyols; about 10-50% by weight of PEG-400; andabout 0.005-0.05% by weight of surfactants.

The excipient may comprise a surfactant. The saccharide may includefructose, trehalose, sorbose, lactose and glucose. The polyol mayinclude mannitol. The surfactant may include polyoxyethylene sorbitanmonooleate (tween 80), poloxamer 188, polyethylene glycol (35), castoroil hydrogenated (RH-35), polyethylene glycol (40), castor oilhydrogenated (RH-40), and polyglycol 12 hydroxystearate.

The mixture solution of the vancomycin hydrochloride solution and theexcipient may comprise: about 25% by weight trehalose; and about 0.01%by weight of polyoxyethylene sorbitan monooleate (tween 80).

The method of filtering the vancomycin concentrate to form the finalfiltrate may comprise: passing the vancomycin concentrate through atleast one of a reverse osmosis membrane and a 200-800 Da nano-filtrationmembrane; and controlling a temperature during filtering so that it doesnot exceed 20° C.

The vancomycin concentrate may have a pH of about 2.0-4.0 and may bestored at a temperature of about 2-8° C. for not more than about 120hours.

The method of spray drying may comprise: turning on a spray dryer,including a heating device; controlling an inlet temperature and anoutlet temperature; turning on a feeding switch; maintaining the spraydryer in a positive pressure; and using above 95% nitrogen as a heatinggas, wherein the inlet temperature is controlled to about 160° C. toabout 240° C. and the outlet temperature is controlled to about 80° C.to about 120° C.

The spray dried vancomycin hydrochloride powder may be sterile.

The method may further comprise: filling the spray dried vancomycinhydrochloride powder in a dosage form; and sealing the dosage whilevacuuminzing and injecting nitrogen gas into the vial to provide asealed dosage form.

The sealed dosage form may comprise a dosage strength of about 0.5 g,about 0.75 g, about 1.0 g, about 2 g, about 5 g, about 10 g or about 20g.

The sterile vancomycin hydrochloride powder may be administrable orallyor injectable intravenously.

The spray dried vancomycin hydrochloride powder may have water contentbelow 3%.

The dosage form may comprise a vial or a capsule.

According to a further aspect of the disclosure, a method is providedfor preparing spray dried powder containing vancomycin hydrochloridethat comprises: providing a vancomycin hydrochloride solution with achromatographic purity of at least 95%; adding a stabilizer and asolubilizer to the vancomycin hydrochloride solution to form a mixturesolution of the vancomycin hydrochloride solution, the stabilizer andthe solubilizer; concentrating the mixture solution to form a 20% to 30%vancomycin concentrate; and spray drying the vancomycin concentrate toform a spray dried vancomycin hydrochloride powder with EP impurity Blevel of not more than 1.5%.

The method may further comprise filtering the vancomycin concentrate toform a final filtrate.

According to a still further aspect of the disclosure, a system isprovided for preparing spray dried powder containing vancomycinhydrochloride. The system comprises: a tank that receives a vancomycinhydrochloride solution with a chromatographic purity of at least 95%; amixer that mixes the vancomycin hydrochloride solution and an excipienthat is added to produce a mixture solution of the vancomycinhydrochloride and the excipient; a membrane that concentrates themixture solution to produce a 20% to 30% vancomycin concentrate; afilter that filters the vancomycin concentrate to form a final filtrate;and a spray dryer that dries the final filtrate to form a spray driedvancomycin hydrochloride powder with EP impurity B level of not morethan 1.5%.

Additional features, advantages, and embodiments of the disclosure maybe set forth or apparent from consideration of the detailed descriptionand drawings. Moreover, it is to be understood that the foregoingsummary of the disclosure and the following detailed description anddrawings are exemplary and intended to provide further explanationwithout limiting the scope of the disclosure as claimed.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which are included to provide a furtherunderstanding of the disclosure, are incorporated in and constitute apart of this specification, illustrate embodiments of the disclosure andtogether with the detailed description serve to explain the principlesof the disclosure. No attempt is made to show structural details of thedisclosure in more detail than may be necessary for a fundamentalunderstanding of the disclosure and the various ways in which it may bepracticed. In the drawings:

FIG. 1 shows a first isomeric form of desamidovancomycin;

FIG. 2 shows a second isomeric form of desmidovancomycin;

FIG. 3 shows two pathways for conversion from vancomycin to succinimide,and for conversion from succinimide to desamidovancomycin;

FIG. 4 shows an example of a system for producing high purity and highpotency vancomycin hydrochloride, according to the principles of thedisclosure;

FIG. 5 shows an example of a process for producing high purity and highpotency vancomycin hydrochloride, according to the principles of thedisclosure; and

FIG. 6 shows a vancomycin B versus time diagram that illustrates astability study of spray dried powder with different excipients.

The present disclosure is further described in the detailed descriptionthat follows.

DETAILED DESCRIPTION OF THE DISCLOSURE

The disclosure and the various features and advantageous details thereofare explained more fully with reference to the non-limiting embodimentsand examples that are described and/or illustrated in the accompanyingdrawings and detailed in the following description. It should be notedthat the features illustrated in the drawings are not necessarily drawnto scale, and features of one embodiment may be employed with otherembodiments as the skilled artisan would recognize, even if notexplicitly stated herein. Descriptions of well-known components andprocessing techniques may be omitted so as to not unnecessarily obscurethe embodiments of the disclosure. The examples used herein are intendedmerely to facilitate an understanding of ways in which the disclosuremay be practiced and to further enable those of skill in the art topractice the embodiments of the disclosure. Accordingly, the examplesand embodiments herein should not be construed as limiting the scope ofthe disclosure. Moreover, it is noted that like reference numeralsrepresent similar parts throughout the several views of the drawings.

According to an aspect of the disclosure, a highly-efficient industrialsystem and method are provided herein for producing high potency andhigh purity vancomycin hydrochloride. The method includes the additionof excipients at an optimal point in the preparation process to avancomycin hydrochloride solution. Excipients used in the method mayinclude stabilizers and/or solubilizers. For instance, the excipientsmay include carbohydrates, polyols, amino acids, organic amines, salts,polymers and surfactants, of which carbohydrates and polyols may be usedas non-specific stabilizers. When using carbohydrates as stabilizers, itmay be desirable to avoid reducing carbohydrates. Sucrose and trehaloseare good choices. Mannitol may be used as a stabilizer and asolubilizer. Trehalose is a stable reducing sugar composed of twoglucose molecules constructed by a,a,1,1-glucosidic bond that mayprovide good protection for organisms because trehalose will form aprotection membrane on the cell surface at heavy conditions such ashigher or lower temperature, high osmotic pressure and dryness toprotect protein molecules from denaturation and deactivation to maintainthe life cycle and bio-characteristics of organisms. Many of the speciesshowing extraordinary resistance to adverse external conditions containa lot of trehalose in their organisms. However, this similar effect hasnot been observed for the other natural carbohydrates, such as, e.g.,sucrose and glucose. This distinct characteristic of trehalose makes ita good activity-protective agent of protein pharmaceuticals, enzymes,vaccines and other biological products.

Amino acids can be used to improve the stability of preparations,especially for some protein preparations. The combination of histidine,glycine, sodium aspartate, glutamine, lysine hydrochloride and 5%mannitol in 10 mM phosphate buffer (pH7.0) can be used to inhibit theaggregate forming of keratinocyte growth factor (KGF).

According to an aspect of the disclosure, excipients are added in thespray drying process to improve stability and to improve productquality, even under severe drying conditions.

Further, an appropriate solubilizer should be added to the vancomycinhydrochloride in the spray drying process, since spray dried vancomycintends to have smaller particle sizes and the reconstitution time tendsto be longer than that of the loose-structured lyophilized product.

Active substances may be fixed on an amorphous matrix, which has a highviscosity environment, to ensure low molecular mobility and reactionactivity. Therefore, the chosen excipient should perform well inembedding or facilitating the embedment of the active substance in theamorphous matrix, as well as improving its glass temperature.Compatibility of active substances, effects on particle forming, anddispersity and flowability should be considered, as will be understoodand appreciated by those having ordinary skill in the art.

FIG. 4 shows an example of a system 100 for producing high potency andhigh purity vancomycin hydrochloride powder that may be filled directlyinto, e.g., a vial. The system 100 includes a tank 170, a membrane 175,a filter 180, and a spray dryer 185. The system 100 may comprise acontainer 190. The system 100 may comprise a fermenter 110, a filter115, a washer/eluter 120, a filter 125, a mixer/tank 130, a separator140, a dissolver 145, a filter 150, a column 155, a filter 160, and/or astorage 165.

According to an aspect of the disclosure, the high purity sterilevancomycin hydrochloride powder produced by the system 100 may betransferred into, e.g., single use bags (not shown). The single use bagsmay be sterilized using gamma irradiation, or other mechanism thatsufficiently sterilizes the bags.

Alternatively, the high purity sterile vancomycin hydrochloride powderproduced by the system 100 may be transferred into an immediate bulkcontainer via an isolator, as is known in the art.

In the fermenter 110, a fermentation process of Amycolatopsis Oriertalisis carried out on an appropriate media containing carbon, nitrogen andinorganic salts at appropriate cultivation conditions. The fermenter 110may produce a vancomycin broth, which may undergo a series ofpurification operations to get a concentrate with a chromatographicpurity of vancomycin above 95%. By adding excipients, the concentrationof vancomycin hydrochloride can be increased to 20-30%. Then, theresultant solution may be spray dried to get a sterile (or non-sterile)vancomycin hydrochloride drug substance, which may be filled underaseptic conditions to get preparations.

Referring to FIG. 4, Amycolatopsis Oriertalis, SIP143491, may beintroduced into the fermenter 110, e.g., as described in Chinese PatentApplication No. CN01132048.6. The cultivation may start from inoculumpreparation, then 1st stage seeding and 2nd stage seeding expansionuntil fermentation at a temperature of about 24-34° C. and a pressure ofabout 0.01-0.08 Mpa for 4-6 days to get a vancomycin broth. Oxygendissolving and pH may be controlled during fermentation. The pH of thevancomycin broth may be adjusted to about 8.5-10.5 with, e.g, sodiumhydroxide solution, as described in Patent 200710198599.4. Thevancomycin broth may be delivered to, and passed through the filter 115to get a clear filtrate.

The filter 115 may include, e.g., a 0.02-0.5 μm ceramic membrane, asdescribed in Patent 200710198599.4 to filter the vancomycin broth into aclear filtrate. The clear filtrate is forwarded to the washer/eluter120.

The washer/eluter 120 may include, e.g., a macroporous resin. The clearfiltrate from the filter 115 is passed through the macroporous resin andthe vancomycin may be eluted down with an acid solution containingsolvent after water washing. The eluent include, e.g., a hydrochloricacid solution containing ethanol. An appropriate amount of activatedcarbon may be added into the collected eluate for discoloring beforeforwarding the eluate to the filter 125 to obtain vancomycin solutionwith a concentration of about 100 mg/mL.

After concentration at the filter 125, the concentrate may beprecipitated and about 6% to about 10% (W/V) of NH₄HCO₃ may be added tothe vancomycin solution in the mixer/tank 130. The pH of the vancomycinsolution may be adjusted to about 7.5-8.5 with a base, such as, e.g.,ammonia. The solution may be stirred for about 45-60 minutes while thevancomycin precipitates from the solution. The precipitation temperatureshould be controlled to about 10-20° C. The solution may be kept stillfor about 16±2 hours and then separated to obtain solid vancomycin basefrom the solution.

The vancomycin solution may be provided to the separator 140 to separatethe vancomycin base from the solution. The vancomycin base may be washedwith ethanol to replace the residual solution and get vancomycin crude.The chromatographic purity of the resultant vancomycin base is not lessthan 80% (HPLC).

The vancomycin crude may be dissolved with purified water in a dissolver145, e.g., as described in Patent 200710187300.5, to get a vancomycinsolution. The dissolved vancomycin solution may be passed through afilter 150 to obtain a clear filtrate. The filter 150 may include, e.g.,a 0.01-0.5 μm ceramic membrane.

The filtrate from the filter 150 may be delivered to and passed throughan ion-exchange resin column 155 to get a qualified eluate with about95% vancomycin B. The ion-exchange resin in the column 155 may include,e.g., a cation ion exchange sephadex gel (Sephadex), a sepharose gel(Sepharose), or the like. Vancomycin filtrate is loaded in the column155 in acid condition, while it is eluted down in base condition byadding, e.g., a basic metal salt, an ammonium salt, or the like.Normally, NH₄+ and Na+ salts are used such as NaCl, NH₄HCO₃, (NH₄)₂CO₃,and the like.

Eluate fractions with above 93% vancomycin B are collected for combiningto obtain the mixed qualified eluate with about 95% vancomycin B(according to HPLC method in USP monograph). Then, the qualified eluateis forwarded to the filter 160 where it may undergo ultra-filtration andnano-filtration to get 12-18% (or higher) vancomycin hydrochlorideconcentrate (a vancomycin solution with a chromatographic purity of notless than 95%). The filtered concentrate may be placed into to storage(e.g., a container, a tank, or the like) and stored at 2-8° C. for apredetermined time (e.g., 48 hours, 72 hours, or the like), but lessthan, e.g., 120 hours.

FIG. 5 shows an example of a process 300 for producing high purity andhigh potency vancomycin hydrochloride, according to the principles ofthe disclosure.

Referring to FIGS. 4 and 5, the vancomycin hydrochloride concentratewith a chromatographic purity of not less than 95% is received in thetank 170 (Step 310). The concentrate may be received from the storage165, or directly from the filter 160. Stabilizers and/or solubilizers172 are added to the vancomycin hydrochloride solution to get a mixturesolution of vancomycin hydrochloride and excipients (Step 320). Themixture solution may be stirred in the tank 170 by a mixer (not shown)(Step 330). The stabilizers may comprise saccharides and/or polyols. Thesolubilizers may comprise polyethylene glycol-400 (PEG-400) and/orsurfactants. The amount of saccharides added to the mixture solution maybe, e.g., about 5-35% by weight of dry vancomycin hydrochloride. Theconcentration of polyols may be, e.g., about 10-50% by weight of dryvancomycin hydrochloride. The concentration of PEG-400 may be, e.g.,about 10-50% by weight of dry vancomycin hydrochloride. The amount ofsurfactants may be, e.g., about 0.005-0.05% by weight of dry vancomycinhydrochloride.

The saccharide may include, e.g., fructose, trehalose, sorbose, lactose,glucose, or the like. The polyol may include, e.g., mannitol, or thelike. The surfactant may include, e.g., polyoxyethylene sorbitanmonooleate (tween 80), poloxamer 188, polyethylene glycol (35), castoroil hydrogenated (RH-35), polyethylene glycol (40), castor oilhydrogenated (RH-40), polyglycol 12 hydroxystearate, or the like.

The amount of trehalose added in the mixture solution may be, e.g.,about 25% by weight, of dry vancomycin hydrochloride.

The amount of polyoxyethylene sorbitan monooleate (tween 80) added tothe mixture solution may be, e.g., about 0.01% by weight of dryvancomycin hydrochloride.

The mixture solution may be concentrated in the membrane equipment 175to obtain 20-30% vancomycin concentrate (Step 340). The membraneequipment 175 may include, e.g., a reverse osmosis membrane. Thetemperature during concentration (e.g., in the membrane equipment 175)may be controlled so as not to exceed 20° C. The vancomycin concentratemay be delivered to the filter 180 for filtration.

The filter 180 receives and filters the vancomycin concentration,outputting a final filtrate (Step 350). The filter 180 may include,e.g., a 200-800 Da nano-filtration member. The temperature duringfiltration may be controlled so as not to exceed 20° C. The finalfiltrate may be delivered to a spray dryer 185 to get spray driedvancomycin hydrochloride powder with EP impurity B level of not morethan 1.5% (Step 360). The pH of the spray dried vancomycin hydrochloridepowder may be, e.g., about 2.0-4.0. The spray dried vancomycinhydrochloride powder may have water content that is below 3%.

The spray dryer 185 may comprise an atomizer (not shown), a feedstockpump (not shown), a gas heater (not shown), a gas disperser (not shown),and a drying chamber (not shown). The spray dryer 185 may furthercomprise a system for cleaning exhaust gases (not shown) and a systemfor recovering vancomycin hydrochloride powder (not shown) and forinjecting the powder into, e.g., vials. The spray dryer 185 may furthercomprise a cyclone. The spray dryer 185 may comprise, e.g., a BuchiB-290 spray dryer, a nano spray dryer, or the like.

According to an aspect of the disclosure, the spray dryer 185 may beoperated by a method that comprises: (1) turning on the spray dryer 185,including the heating device (not shown); (2) controlling andmaintaining inlet temperature to, e.g., about 160-240° C., andcontrolling and maintaining outlet temperature to, e.g., 80-120° C.; (3)turning on the feeding switch (not shown); (4) maintaining the spraydryer 185 at positive pressure; and (5) using above 95% nitrogen asheating gas.

The vancomycin hydrochloride powder delivered from the spray dryer 185may be sterile or non-sterile. If the spray dried vancomycinhydrochloride powder is sterile, then it may be filled into vials withAuger, e.g., using an aseptic filling machine (not shown), and thencapped while vacuumizing and filling nitrogen. The dosage strength mayinclude, e.g., about 0.5 g, 0.75 g, 1.0 g, 2 g, 5 g, 10 g, or 20 g.

Further, sterile vancomycin hydrochloride powder may be administered byoral or injectable form.

Preferably, the vancomycin hydrochloride solution with a chromatographicpurity of not less than 95% is prepared by dissolving vancomycinhydrochloride active pharmaceutical ingredient (API) in purified water,or in water for injection.

Preferably, the amount of saccharides is 5-35% by weight of dryvancomycin hydrochloride, the concentration of polyols is 10-50 wt. %and the concentration of PEG-400 is 10-50 wt. %, the amount ofsurfactants is 0.005-0.05% by weight of dry vancomycin hydrochloride.

Preferably, the saccharide includes trehalose, fructose, sorbose,lactose and glucose, the polyol includes mannitol, the surfactantincludes polyoxyethylene sorbitan monooleate (tween 80), poloxamer 188,polyethylene glycol (35), castor oil hydrogenated (RH-35), polyethyleneglycol (40), castor oil hydrogenated (RH-40), and polyglycol 12hydroxystearate.

Preferably, the amount of trehalose is 25% by weight of dry vancomycinhydrochloride, and the amount of polyoxyethylene sorbitan monooleate(tween 80) is 0.01% by weight of dry vancomycin hydrochloride.

Preferably, the membrane concentration equipment 175 (shown in FIG. 4)includes a reverse osmosis membrane and a 200-400 Da nano-filtrationmembrane. The temperature during concentration is controlled not morethan 20° C.

Preferably, the pH of the final concentrate is 2.0-4.0, and theconcentrate is stored at 2-8° C. for not more than 120 hours.

Preferably, for the spray drying operation (Step 360), the operationincludes turning on the spray dryer 170 and heating device (not shown,in spray dryer 170, FIG. 4), controlling inlet temperature and outlettemperature at 160-240° C. and 80-120° C., respectively, turning onfeeding switch (not shown, in spray dryer 170), keeping the system inpositive pressure and using above 95% nitrogen as heating gas.

Preferably, the spray dried vancomycin hydrochloride powder can besterile or non-sterile.

Preferably, if the spray dried vancomycin hydrochloride powder issterile, it can be filled in vials using, e.g., an aseptic fillingmachine with Auger, and then capped while vacuumizing and fillingnitrogen. The strengths are 0.5 g, 0.75 g, 1.0 g, 2 g, 5 g, 10 g and 20g.

Preferably, the sterile vancomycin hydrochloride can be administered byoral or injectable form.

Preferably, the spray dried vancomycin hydrochloride powder has watercontent below 3%.

In the system 100 and/or process 300, by adding an appropriate ratio ofexcipients to a vancomycin solution at the optimal point in the process,as disclosed herein, the vancomycin hydrochloride solution is stabilizedand the concentration of vancomycin is increased to 20-30%, whilesimultaneously increasing drying efficiency. Further, the instantdisclosure can be implemented to provide high concentration, sterilevancomycin hydrochloride drug substances with favorable bulk density,which allow for easy filling of vials. The optimally added excipientsimprove product stability in spray drying processes; reduce the numberof process steps; control and maintain low impurity levels; and improveproduct solubility and quality.

The present disclosure realizes commercial production of high qualityand high potency vancomycin hydrochloride from fermentation topreparation and filling of sterile drug substances, then to sterilepowder for injection after filling. Compared to existing technology, theproduction process disclosed herein shortens production time, has fewerproduction steps, reduces cost, and improves yield, allowing forimplementation in industrial scale production.

The following examples are presented to illustrate the presentdisclosure. The disclosure is not to be considered limited by theseexamples nor the parameter ranges in these examples.

EXAMPLES Example 1 Stability Study of Vancomycin Concentrates withDifferent Excipients

To 900 mL of 25% vancomycin concentrate (batch No: 310121109; vancomycinB: 96.50%; 225 g of vancomycin in total), subdivided in nonuplicate, 100mL each. To eight of them, added 5 g of trehalose, 5 g of mannitol, 5 gof glucose, 5 g of fructose, 5 g of glycine, 5 g of glutamine, 5 g ofsorbose and 5 g of lactose, respectively, passed these eight solutionsas well as the one without excipients through 0.22 μm filter,respectively, and stored the obtained filtrate at 2-8° C. inrefrigerator. Observed clarity of the filtrate every 24 hours and theresults are shown in Table 1.

TABLE 1 Clarity of vancomycin concentrate at 2-8° C. with differentamounts of trehalose Solution Solution Solution Solution Amount ofappearance after appearance after appearance after appearance afterexcipient (g) 24 hours 48 hours 72 hours 96 hours 0 White particle Whiteparticle White particle Colloidal, precipitate precipitate precipitatewithout fluidity 5 g trehalose Clear Clear Clear Clear 5 g mannitolClear Clear Clear Filamentous precipitate 5 g glucose Clear Clear Whiteparticle White particle precipitate precipitate 5 g fructose Clear ClearWhite particle White particle precipitate precipitate 5 g glycine ClearWhite particle White particle White particle precipitate precipitateprecipitate 5 g glutamine Clear White particle White particle Colloidal,precipitate precipitate without fluidity 5 g sorbose Clear Clear Whiteparticle White particle precipitate precipitate 5 g lactose Clear ClearWhite particle White particle precipitate precipitate

Based on the data in Table 1, it can be seen that appropriate excipientscan improve the stability of vancomycin concentrate at 2-8° C., of whichtrehalose is the best one.

Example 2 Effect of Different Trehalose Concentration on Stability ofVancomycin Concentrate

To 800 mL of 25% vancomycin concentrate (batch No: 310121109; vancomycinB: 96.50%; 200 g of vancomycin in total), subdivided in octuplicate, 100mL (about 25 g of vancomycin) each.

To the eight portions, add 0 g, 1.25 g (5% of weight of vancomycin), 2.5g (10% of weight of vancomycin), 3.75 g (15% of weight of vancomycin), 5g (20% of weight of vancomycin), 6.25 g (25% of weight of vancomycin),7.5 g (30% of weight of vancomycin) and 8.75 g (35% of weight ofvancomycin) of trehalose, respectively, dissolved completely andfiltered through 0.22 μm to get Filtrate A, B, C, D, E, F, G and H,respectively. Stored all the obtained filtrates at 2-8° C. in arefrigerator, and observed clarity of the filtrates every 24 hours. Theresults are presented in Table 2.

TABLE 2 Clarity of vancomycin concentrate at 2-8° C. with differentamount of trehalose Solution Solution Solution Solution SolutionSolution Amount appearance appearance appearance appearance appearanceappearance of after 24 after 48 after 72 after 96 after 120 after 144trehalose hours hours hours hours hours hours 0 White White WhiteColloidal, Colloidal, Colloidal, particle particle particle withoutwithout without precipitate precipitate precipitate fluidity fluidityfluidity  5% Clear Clear Clear Clear Filamentous Filamentous precipitateprecipitate 10% Clear Clear Clear Clear Clear Filamentous precipitate15% Clear Clear Clear Clear Clear Clear 20% Clear Clear Clear ClearClear Clear 25% Clear Clear Clear Clear Clear Clear 30% Clear ClearClear Clear Clear Clear 35% Clear Clear Clear Clear Clear Clear

Based on the data in Table 2, it can be seen that vancomycin concentrateis still clear after storage at 2-8° C. for 144 hours when the amount ofadded trehalose is 15%-35% by weight of vancomycin.

Example 3 Process Parameters Study on Spray Drying of VancomycinConcentrate

Spray dryer: Buchi B-290.

Materials: vancomycin concentrate, batch No: 310121109; vancomycin B:96.50%; Concentration: 20%.

Experiment process: in these embodiments, the spray drying was performedat three inlet temperatures of 170° C., 190° C. and 210° C., and foreach inlet temperature, three outlet temperature ranges of 90-100° C.,100-110° C. and 110-120° C. were adopted, respectively.

For each embodiment, at the chosen inlet air temperature and thecorresponding outlet air temperature by adjusting speed of feedstockpump, performed spray drying of 200 mL vancomycin concentrate, collecteddry vancomycin powder and tested moisture, pH of 5% vancomycin solution,HPLC as well as OD of 10% vancomycin solution.

The batch numbers of spray dried vancomycin hydrochloride powdertogether with the corresponding inlet temperature and outlet temperatureare listed as follows:

Inlet Outlet Batch. No of temperature temperature spray dried (° C.) (°C.) powder 170° C.  90-100° C. 112001 100-110° C. 112002 110-120° C.112003 190° C.  90-100° C. 112004 100-110° C. 112101 110-120° C. 112102210° C.  90-100° C. 112103 100-110° C. 112104 110-120° C. 112105

The experimental records and results are described as follows:

TABLE 3-1 Records of spray drying Test No. 112001 112002 112003 112004Duration 10:45-11:10 14:10-14:50 15:25-16:10 16:30-16:55 InletTemperature (° C.) 170  170  170  190  Outlet Temperature (° C.)  90-100100-110 110-120  90-100 Air volume (%) 100  100  100  100  Speed of pump(%) 30 25 20 40 Flow rate of nitrogen (mm) 40 50 50 50 Systempressure(mbar) 35 15 15 15

TABLE 3-2 Records of spray drying Test No. 312101 112102 112103 112104112105 Duration 8:35-9:05 10:02-10:35 11:05-11:40 15:00-15:3016:10-16:55 Inlet Temperature (° C.) 190  190  210  210  210  OutletTemperature (° C.) 100-110 110-120  90-100 100-110 110-120 Air volume(%) 100  100  100  100  100  Speed of pump (%) 30 25 50 40 30 Flow rateof nitrogen (mm) 40 50 50 40 50 System pressure (mbar) 15 15 15 15 15

TABLE 4-1 Test results of spray dried powder Original Test No. test112001 112002 112003 112004 Water (%) NA 6.2 5.9 6.7 5.9 pH, 5% Conc.2.93 3.08 3.08 3.10 3.07 A_(450,) 10% Conc. 0.054 0.068 0.064 0.0740.069 Assay of 96.5 95.44 95.22 95.49 95.47 vancomycin B (%) Impurity B₁0.79 0.89 0.90 0.89 0.88 Impurity B₂ / 0.14 0.18 0.14 0.14 Impurity B₁and B₂ 0.79 1.03 1.08 1.03 1.02

TABLE 4-2 Test results of spray dried powder Test No. 112101 112102112103 112104 112105 Water (%) 5.8 6.2 5.7 6.1 5.5 pH, 5% Conc. 3.053.08 3.01 3.02 3.05 A_(450,) 10% Conc. 0.067 0.067 0.062 0.062 0.075Assay of 95.34 95.24 95.13 95.14 95.02 vancomycin B (%) Impurity B₁ 0.940.94 0.98 0.95 0.96 Impurity B₂ 0.19 0.18 0.19 0.16 0.25 Impurity B₁ andB₂ 1.13 1.12 1.17 1.11 1.21

Referring to the data in Tables 4-1 and 4-2, it can be seen that: (1)without addition of excipient, vancomycin B in the final dried powderdecreased, at the same time, impurity B₁ and impurity B₂ increased; (2)the dried powders obtained at inlet air temperature of 170° C., 190° C.and 210° C. have equivalent quality, and the same for outlet airtemperature of 90-100° C., 100-110° C. and 110-120° C.; and (3) all thespray dried powder has higher water content.

Example 4 Spray Drying Comparison of Vancomycin Concentrate of DifferentConcentrations with 25% Trehalose

To 1600 mL of vancomycin concentrate (Batch No: 310121110, vancomycin B:96.28%) with a vancomycin concentration of 15% (containing 240 g ofvancomycin in total), added 60 g of trehalose (25% by weight ofvancomycin), mixed well and concentrated through D200 nano-filtrationmembrane to 800 mL (30% vancomycin concentration), passed through 0.22μm filter, then subdivided the obtained filtrate in quadruplicate, 200mL each.

To these four portions, added 0 mL, 40 mL, 100 mL and 200 mL of purifiedwater, respectively, mixed well to get solution A (30% vancomycin), B(25% vancomycin), C (20% vancomycin) and D (15% vancomycin).

Spray drying of solution A, B, C and D was performed using a drying gashaving an inlet temperature of 190° C. and an outlet temperature of100-110° C. Resultant powders were tested as shown in Table 6.

TABLE 5 Test results of spray dried powders of vancomycin concentrate atdifferent concentrations Test No. 30% Conc. 25% Conc. 20% Conc. 15%Conc. Duration  9:35-10:02 10:30-11:10 14:05-14:53 13:37-16:45 InletTemperature (° C.) 190  190  190  190  Outlet Temperature (° C.) 100-110100-110 100-110 100-110 Air volume (%) 100  100  100  100  Speed of pump(%) 35 30 25 20 Flow rate of nitrogen (mm) 50 50 50 50 System pressure(mbar) 15 15 15 15

TABLE 6 Test results of spray dried powder of different vancomycinconcentrations Original 30% 25% 20% 15% Test No. test Conc. Conc. Conc.Conc. Moisture (%) NA 3.8 3.7 4.1 4.3 pH, 5% Conc. 2.81 2.89 2.81 2.812.82 A_(450,) 10% Conc. 0.054 0.057 0.057 0.052 0.052 Vancomycin B (%)96.28 96.24 96.22 96.18 96.15 Impurity B₁ (%) 0.96 0.94 0.95 0.95 0.96Impurity B₂ (%) 0.05 0.06 0.08 0.06 0.07 Impurity B₁ and B₂ (%) 1.011.00 1.03 1.01 1.03 * Impurity B1: desamidovancomycin 1; Impurity B2:desamidovancomycin 2.

The results showed no significant quality difference for the spray driedvancomycin powders at different vancomycin concentrations. However, ahigher vancomycin concentration can reduce drying time, reduce energycosts and increase efficiency.

Example 5 Spray Drying Study of 25% Vancomycin Concentrate withDifferent Amounts of Trehalose

To 3000 mL of vancomycin concentrate (Batch No: 310121113, vancomycin B:96.28%) with a vancomycin concentration of 10% (containing 300 g ofvancomycin in total), subdivided the obtained filtrate in sextuplicate,500 mL each (50 g of vancomycin each).

To these six portions, added 2.5 g (5% by weight of vancomycin), 5 g(10% by weight of vancomycin), 7.5 g (15% by weight of vancomycin), 10 g(20% by weight of vancomycin), 12.5 g (25% by weight of vancomycin) and15 g (30% by weight of vancomycin) of trehalose, respectively, mixedwell and concentrated through D200 nano-filtration membrane to 200 mL(25% vancomycin), then passed through 0.22 μm filter, collected thefiltrates and marked as filtrate A, B, C, D, E and F, respectively.

Spray drying of filtrate A, B, C, D, E and F was performed using adrying gas having an inlet temperature of 190° C. and an outlettemperature of 100-110° C. The results of the experiments are shownbelow in Table 7 and 8.

TABLE 7 Spray drying data of vancomycin concentrate with differentamount of trehalose Trehalose Trehalose Trehalose Trehalose TrehaloseTrehalose 5% 10% 15% 20% 25% 30% Test No. (W/W) (W/W) (W/W) (W/W) (W/W)(W/W) Duration 8:33-9:12  9:30-10:04 10:36-11:17 13:32-14:05 14:35-14:5915:30-16:01 Start-stop Inlet Temperature, ° C. 190 190 190 190 190 190Outlet 100-110 100-110 100-110 100-110 100-110 100-110 Temperature, ° C.Air volume (%) 100 100 100 100 100 100 Speed of pump (%) 20 25 25 30 3035 Flow rate of 50 50 50 50 50 50 Nitrogen (mm) System pressure 15 15 1515 15 15 (mbar)

TABLE 8 Test results of spray dried powder of vancomycin concentrationwith different amount of trehalose Trehalose Trehalose TrehaloseTrehalose Trehalose Trehalose 5% 10% 15% 20% 25% 30% Test No.Concentrate (W/W) (W/W) (W/W) (W/W) (W/W) (W/W) Moisture, % NA 4.2 3.73.4 3.5 3.8 3.6 pH, 5% Conc. 2.81 2.89 2.81 2.89 2.84 2.85 2.81 A_(450,)10% Conc. 0.054 0.052 0.057 0.057 0.052 0.052 0.055 Vancomycin B 96.2896.09 96.14 96.13 96.24 96.12 96.22 (%) Impurity B₁ (%) 0.96 0.93 0.940.94 0.94 0.94 0.95 Impurity B₂ (%) / 0.11 0.08 0.06 0.06 0.05 0.03Impurity B₁ and 0.96 1.04 1.02 1.00 1.00 0.96 0.98 B₂ (%)

From the data provided in Table 8, it can be seen that the spray driedpowder has a similar quality when the amount of trehalose is from 5% to30%.

Example 6 Spray Drying Study of Vancomycin Concentrate with DifferentAmounts of Tween 80

To 2500 mL of vancomycin concentrate (Batch No: 310130102, vancomycin B:96.08%) with a vancomycin concentration of 10% (containing 250 g ofvancomycin), added 62.5 g of trehalose (25% by weight of vancomycin),concentrated through D200 nano-filtration membrane to 1000 mL with aconcentration of 25%, subdivided solution in quintuplicate, 200 mL each(50 g of vancomycin each).

For these five portions, added 0 mL, 0.1 mL (0.005% by weight ofvancomycin), 0.2 mL (0.01% by weight of vancomycin), 0.4 mL (0.02% byweight of vancomycin) and 1 mL (0.05% by weight of vancomycin) of 2.5%Tween 80 solution, respectively, mixed well and passed through 0.22 μmfilter to obtain filtrate A, B, C, D and E, respectively.

Spray drying of filtrate A, B, C, D and E was performed using a dryinggas having an inlet temperature of 190° C. and an outlet temperature of100-110° C. The results of the experiments are shown below in Table 9and 10.

TABLE 9 Spray drying data of vancomycin concentrate with differentamount of Tween-80 Tween 80 Tween 80 Tween 80 Tween 80 Tween 80 Test No.0.00% 0.05% 0.02% 0.01% 0.005% Duration Start-stop 9:03-9:32  9:55-10:3410:57-11:23 13:45-14:15 14:55-13:29 Inlet Temperature, ° C. 190  190 190  190  190  Outlet Temperature, ° C. 100-110 100-110 100-110 100-110100-110 Air volume (%) 100  100  100  100  100  Speed of pump (%) 20 2525 30 30 Flow rate of Nitrogen (mm) 50 50 50 50 50 System pressure(mbar) 15 15 15 15 15

TABLE 10 Test results of spray dried vancomycin powder with differentamount of Tween-80 Tween 80 Tween 80 Tween 80 Tween 80 Tween 80 Test No.Concentrate 0.00% 0.05% 0.02% 0.01% 0.005% Moisture % NA 4.2 2.1 2.3 2.52.8 pH, 5% Conc. 2.85 2.89 2.84 2.87 2.85 2.85 A₄₅₀, 10% Conc. 0.0520.057 0.057 0.056 0.055 0.053 Vancomycin B (%) 96.08 96.01 96.02 96.0396.04 96.02 Impurity B₁ (%) 0.69 0.70 0.70 0.69 0.68 0.70 Impurity B₂(%) / 0.05 0.03 0.06 / / Impurity B₁ and B₂ 0.69 0.75 0.73 0.75 0.680.70 (%) Reconstitution Time NA 28 S 15 S 16 S 14 S 17 S (Seconds, 5%Conc.)* *Reconstitution time of 5% Conc. means the required time oftotally dissolving of containing vancomycin hydrochloride 0.5 g samplein 10 mL WFI in 15 ml Vials.

From the data in Table 10, it can be concluded that: (1) the watercontent of spray dried powder decreases significantly when addingtween-80; and (2) the reconstitution time of spray dried powder becomesshort when adding tween-80.

Example 7 Spray Drying Comparison of Vancomycin Concentration withTrehalose and Tween 80 or Mannitol and PEG

Subdivided 2000 mL of concentrate (batch No: 310130105, vancomycin B:96.40%) with a 15% vancomycin concentration (containing 300 g ofvancomycin in total) in duplicate, 150 g vancomycin each. To one ofthem, added 37.5 g of trehalose (25% by weight of vancomycin), dissolvedand concentrated through D200 nano-filtration membrane to 600 mL, thenadded 0.6 mL of 2.5% tween-80 solution (0.01% of by weight vancomycin),mixed well and passed through 0.22 μm filter to get filtrate A. To theother one, added 30 g of mannitol (20% of vancomycin by weight) and 30 gof PEG-400 (20% of vancomycin by weight), mixed well, concentratedthrough D200 nano-filtration membrane to 600 mL, then passed through0.22 μm filter, collected the filtrate and marked as filtrate B.

Subdivided filtrate A in triplicate, 200 mL each. Conducted spray dryingrespectively with an inlet temperature 190° C. and an outlet temperatureof 100-110° C. and the obtained dried powder batches are 0204, 0205 and0206, respectively. Conducted the same operations for filtration B andthe obtained batches are 0201, 0202 and 0203, respectively. The resultsof the experiments are shown below in Table 11 and 12, respectively.

TABLE 11 Spray drying data of vancomycin concentrate with differentformualtions Formulations Formulations Trehalose, 25% (W/W) Mannitol,20% (W/W) Tween 80, 0.01% PEG400, 20% Vanconmycin B, 74.99% VanconmycinB, 60% Test No. 0204 0205 0206 0201 0202 0203 Duration 13:34-14:1214:30-15:04 15:40-16:17  8:34-9:08  9:49-10:20 10:40-11:09 Start-stopInlet Temp., ° C. 190 190 190 190 190 190 Outlet Temp., ° C. 100-110100-110 100-110 100-110 100-110 100-110 Air volume (%) 100 100 100 100100 100 Speed of pump 30 30 30 30 30 30 (%) Flow rate of 50 50 50 50 5050 Nitrogen (mm) System pressure 15 15 15 15 15 15 (mbar)

TABLE 12 Test results of spray dried vancomycin powder with differentformulations Formulations Formulations Trehalose, 25% (W/W) Mannitol,20% (W/W) Tween 80, 0.01% PEG400, 20% Vanconmycin B, 74.99% VanconmycinB, 60% Test No. Concentrate 0204 0205 0204 0205 0204 0205 Moisture % NA2.2 2.4 2.4 2.5 2.8 2.6 pH, 10% Conc. 3.05 3.11 3.14 3.10 3.15 3.13 3.11A₄₅₀, 10% Conc. 0.056 0.052 0.054 0.055 0.062 0.054 0.053 Vancomycin B(%) 96.40 96.16 96.31 96.26 95.86 95.93 95.87 Impurity B₁ (%) 0.55 0.540.52 0.55 0.70 0.71 0.72 Impurity B₂ (%) / 0.07 / / 0.07 0.07 0.07Impurity B₁ and B₂ (%) 0.55 0.61 0.52 0.55 0.77 0.78 0.79 ReconstitutionTime NA 15 17 16 23 25 22 (Seconds, 5% Conc.)* *Reconstitution time of5% Conc. means the required time of totally dissolving of containingvancomycin hydrochloride 0.5 g sample in 10 mL WFI in 15 ml Vials.

From the data in Table 12, it can be conclude that: (1) the spray driedvancomycin powder with the formulation of trehalose and tween 80 hashigher vancomycin B and a lower level of impurity B; and (2) the spraydried vancomycin powder with the formulation of trehalose and tween 80has a shorter reconstitution time at a concentration of 5%.

Example 8 Stability Study of Spray Dried Vancomycin Powder with andWithout Excipients

Subdivided 1500 mL (containing 150 g of vancomycin in total) ofconcentrate (batch No: 310130208, vancomycin B: 96.40%) with a 10%vancomycin concentration in triplicate, 50 g of vancomycin and 500 mLeach.

To one of them, added 12.5 g of trehalose (25% by weight of vancomycin),dissolved and concentrated through D200 nano-filtration membrane to 200mL, then added 0.5 mL of 1.0% tween-80 solution (0.01% by weight ofvancomycin), mixed well and passed through 0.22 μm filter to getfiltrate A with a vancomycin concentration of 25%. Concentrated theother one through D200 nano-filtration membrane to 200 mL, then passedthrough 0.22 μm filter to get filtrate B with a vancomycin concentrationof 25%. For the last one, added 10 g of mannitol (20% by weight ofvancomycin), and 10 g of PEG-400 (20% by weight of vancomycin),dissolved and concentrated through D200 nano-filtration membrane, thenpassed through 0.22 μm filter to get filtrate C with a vancomycinconcentration of 25%.

Conducted spray drying respectively with an inlet temperature 190° C.and an outlet temperature of 100-110° C. The results of the experimentsare shown below in Tables 13 and 14, respectively.

Stored the spray dried powder in 60° C. chamber and tested the impuritylevel by HPLC periodically. The results are shown in Table 15.

TABLE 13 Spray drying data of vancomycin concentrate with and withoutexcipients Formulations Trehalose, Vanconmycin Mannitol Tween 80 WithoutPEG-400 Test No. Vanconmycin excipients Vancomycin Duration Start-stop8:55-9:10  9:40-10:18 10:40-11:18 Inlet Temperature, ° C. 190  190  190 Outlet Temperature, ° C. 100-110 100-110 100-110 Air flow (%) 100  100 100  Speed of pump (%) 30 25 30 Flow rate of 50 50 50 Nitrogen (mm)System pressure (mbar) 15 15 15

TABLE 14 Product results of spray dried vancomycin powder with andwithout excipients Trehalose, Vanconmycin Mannitol Tween 80 WithoutPEG-400 Test No. Vanconmycin excipients Vancomycin Moisture, % 2.8 4.72.7 pH, 5% Conc. 2.95 2.91 2.91 A₄₅₀, 10% Conc. 0.052 0.057 0.053Vancomycin B (%) 96.16 95.58 95.80 Impurity B₁ (%) 0.54 0.57 0.70Impurity B₂ (%) 0.07 0.13 0.07 Impurity B₁ and B₂ (%) 0.61 0.70 0.77Reconstitution Time 15 S 24 S 14 S (Seconds, 5% Conc.)* *Reconstitutiontime of 5% Conc. means the required time of totally dissolving ofcontaining vancomycin hydrochloride 0.5 g sample in 10 mL WFI in 15 mlVials.

TABLE 15 Stability study of spray dried vancomycin powder with andwithout excipients Day 1, Day 2, Day 3, Day 5, Test No. 60° C. 60° C.60° C. 60° C. Trehalose and tween 80 Vancomycin B (%) 95.76 95.35 94.9594.48 Impurity B₁ (%) 0.82 0.85 1.08 1.25 Impurity B₂ (%) 0.08 0.17 0.250.29 Impurity B₁ and B₂ (%) 0.90 1.02 1.33 1.54 Without excipientVancomycin B (%) 93.64 92.82 93.36 90.82 Impurity B₁ (%) 0.93 1.18 1.301.76 Impurity B₂ (%) 0.46 0.64 0.71 1.00 Impurity B₁ and B₂ (%) 1.391.82 2.01 2.76 Mannitol and PEG Vancomycin B (%) 95.08 94.48 93.94 93.31Impurity B₁ (%) 0.88 1.33 1.37 1.73 Impurity B₂ (%) 0.11 0.16 0.17 0.20Impurity B₁ and B₂ (%) 0.99 1.49 1.54 1.93

FIG. 6 shows a vancomycin B versus time diagram that illustrates theresults of the stability study of the spray dried powder with differentexcipients. In the diagram, curve 1 denotes spray dried powder withtrehalose and tween 80, curve 2 denotes spray dried powder withoutexcipient, and curve 3 denotes spray dried powder with mannitoal andPEG-400.

From the data in Table 15, it can be seen that the formulation withtrehalose and tween has the best product stability, followed by theformulation of mannitol and PEG. Vancomycin without excipients has theworst stability.

Example 9 Spray Drying Study of Vancomycin Concentrate at DifferentConcentrations

To 1800 mL (containing 180 g of vancomycin in total) of vancomycinconcentrate (batch No: 310130210, vancomycin B: 96.05%) with a 10%vancomycin concentration, added 45 g of trehalose (25% by weight ofvancomycin), dissolved and concentrated through D200 nano-filtrationmembrane to 600 mL, then added 1.8 mL of 1.0% tween-80 solution (0.01%by weight of vancomycin), mixed well and passed through 0.22 μm filter.Subdivided the obtained filtrate in triplicate, 200 mL each (containing60 g of vancomycin each). Added 40 mL, 100 mL and 0 mL of purifiedwater, respectively, mixed well to get solution A (25% vancomycin), B(20% vancomycin) and C (30% vancomycin).

Spray drying of solution A, B and C were performed with an inlettemperature of 190° C. and an outlet temperature of 100-110° C. Theresults of the experiments are shown below in Tables 16 and 17,respectively.

TABLE 16 Spray drying data of vancomycin concentrate at differentconcentrations Test No. 30% Conc. 25% Conc. 20% Conc. DurationStart-stop 14:15-14:46 15:10-15:55 16:35-17:35 Inlet Temperature, ° C.190  190  190  Outlet Temperature, ° C. 100-110 100-110 100-110 Airvolume (%) 100  100  100  Speed of pump (%) 30 30 25 Flow rate ofNitrogen (mm) 50 50 50 System pressure (mbar) 15 15 15

TABLE 17 Test results of spray dried vancomycin powder 30% 25% 20% TestNo. Concentrate Conc. Conc. Conc. Moisture, % NA 2.8 2.7 3.1 pH, 5%Conc. 3.05 3.14 3.11 3.16 A₄₅₀, 10% Conc. 0.046 0.052 0.047 0.051Vancomycin B (%) 96.05 96.01 95.91 95.92 Impurity B₁ (%) 0.67 0.63 0.640.64 Impurity B₂ (%) / / / / Impurity B₁ and B₂ (%) 0.67 0.63 0.64 0.64Reconstitution Time NA 17 14 15 (Seconds, 5% Conc.)* *Reconstitutiontime of 5% Conc. means the required time of totally dissolving ofcontaining vancomycin hydrochloride 0.5 g sample in 10 mL WFI in 15 mlVials.

From the data in Table 17, it can be seen that the spray dried powderobtained by vancomycin solutions with different concentrations 20%, 25%and 30% has similar quality.

Example 10 Spray Drying of Vancomycin Concentrate with DifferentFormulations

To 3500 mL (containing 350 g of vancomycin in total) of vancomycinconcentrate (batch No: 310130208, vancomycin B: 96.40%) with a 10%vancomycin concentration, added 87.5 g of trehalose (25% by weight ofvancomycin), dissolved and concentrated through D200 nano-filtrationmembrane to 1400 mL, subdivided the obtained filtrate in septuplicate,200 mL each (containing 50 g of vancomycin each).

To six of them, added 0.5 ml of 1.0% tween-80 solution (0.01% by weightof vancomycin), 2.5 ml of 1.0% poloxamer 188 (0.05% by weight ofvancomycin), 2.5 ml of 1.0% polyethylene glycol (0.05% by weight ofvancomycin), 2.5 ml of 1.0% castor oil hydrogenated (0.05% by weight ofvancomycin), 2.5 ml of 1.0% polyglycol 12 hydroxystearate (0.05% byweight of vancomycin), and 2.5 ml of 1.0% polyoxyethylene castor oilhydrogenated, respectively, mixed well and passed through 0.22 μmfilter, collected the filtrate and marked as filtrate A, C, D, E, F andG, respectively. Each one has a vancomycin concentration of 25%.

To the last one, passed through 0.22 μm filter, collected the filtrateand marked as filtrate B.

Spray drying of solution A, B, C, D, E, F and G were performed with aninlet temperature of 190° C. and an outlet temperature of 100-110° C.The results of the experiments are shown below in Table 18.

TABLE 18 Test results of spray dried vancomycin powder ReconstitutionTime (Seconds, pH, 5% Test No. Moisture, % 5% Conc.)* Conc. Tween-80 2.114 2.92 Blank 3.9 29 2.91 Poloxamer 188 2.8 21 2.93 Polyethylene glycol3.0 19 2.92 Castor oil 3.1 19 2.91 hydrogenated Polyglycol 12 3.2 222.93 hydroxystearate Polyoxyethylene 2.6 18 2.95 castor oil Hydrogenated*Reconstitution time of 5% Conc. means the required time of totallydissolving of containing vancomycin hydrochloride 0.5 g sample in 10 mLWFI in 15 ml Vials.

From the data in table 18, it can be concluded that spray dried powderwith excipients, preferably tween-80, has a faster dissolving rate.

The above examples are meant to be illustrative of the disclosure and donot limit the disclosure. It will be readily apparent to those ofordinary skill in the art that certain changes and modifications may bemade thereto without departing from the spirit or scope of the appendedclaims.

The terms “including,” “comprising” and variations thereof, as used inthis disclosure, mean “including, but not limited to,” unless expresslyspecified otherwise.

The terms “a,” “an,” and “the,” as used in this disclosure, means “oneor more,” unless expressly specified otherwise.

Devices that are in communication with each other need not be incontinuous communication with each other, unless expressly specifiedotherwise. In addition, devices that are in communication with eachother may communicate directly or indirectly through one or moreintermediaries.

Although process steps, method steps, algorithms, or the like, may bedescribed in a sequential order, such processes, methods and algorithmsmay be configured to work in alternate orders. It is noted that allprocess steps disclosed herein may be carried out aseptically. In otherwords, any sequence or order of steps that may be described does notnecessarily indicate a requirement that the steps be performed in thatorder. The steps of the processes, methods or algorithms describedherein may be performed in any order practical. Further, some steps maybe performed simultaneously.

When a single device or article is described herein, it will be readilyapparent that more than one device or article may be used in place of asingle device or article. Similarly, where more than one device orarticle is described herein, it will be readily apparent that a singledevice or article may be used in place of the more than one device orarticle. The functionality or the features of a device may bealternatively embodied by one or more other devices which are notexplicitly described as having such functionality or features.

What is claimed:
 1. A method for preparing a spray dried powdercomprising vancomycin hydrochloride, the method comprising the steps of:providing a vancomycin hydrochloride solution with a chromatographicpurity of at least 95%; adding an excipient to the vancomycinhydrochloride solution to form a mixture solution of the vancomycinhydrochloride solution and the excipient; concentrating the mixturesolution of the vancomycin hydrochloride solution and the excipient toform a concentrate comprising about 20% to 30% (w/v) vancomycinhydrochloride; filtering the vancomycin concentrate through at least oneof a reverse osmosis membrane and a 200-800 Da nano-filtration membraneand controlling a temperature during filtering so that it does notexceed 20° C.; and spray drying the final filtrate to form a spray driedvancomycin hydrochloride powder with EP impurity B level of not morethan 1.5%.
 2. The method of claim 1, wherein the excipient comprises atleast one of: a stabilizer; and a solubilizer.
 3. The method of claim 1,wherein the excipient comprises a polyol.
 4. The method of claim 2,wherein the excipient comprises at least one of: polyethylene glycol-400(PEG-400); and a surfactant.
 5. The method of claim 1, wherein theproviding the vancomycin hydrochloride solution with the chromatographicpurity of at least 95% comprises: dissolving a vancomycin hydrochlorideactive pharmaceutical ingredient (API) in purified water.
 6. The methodof claim 3, the excipient further comprising a surfactant, wherein thepolyol comprises mannitol, and wherein the surfactant is selected fromthe group consisting of polyoxyethylene sorbitan monooleate (tween 80),poloxamer 188, polyethylene glycol (35), castor oil hydrogenated(RH-35), polyethylene glycol (40), castor oil hydrogenated (RH-40), andpolyglycol 12 hydroxystearate.
 7. The method of claim 1, wherein saidvancomycin concentrate has a pH of about 2.0-4.0 and is stored at atemperature of about 2-8° C. for not more than about 120 hours.
 8. Themethod of claim 1, wherein the spray drying comprises: turning on aspray dryer, including a heating device; controlling an inlettemperature and an outlet temperature; turning on a feeding switch;maintaining the spray dryer in a positive pressure; and using above 95%nitrogen as a heating gas, wherein the inlet temperature is controlledto about 160-240° C. and the outlet temperature is controlled to about80-120° C.
 9. The method of claim 1, wherein the spray dried vancomycinhydrochloride powder is sterile.
 10. The method of claim 9, furthercomprising: filling the spray dried vancomycin hydrochloride powder in avial; and sealing the vial while vacuuminizing and injecting nitrogengas into the vial to provide a sealed dosage form.
 11. The method ofclaim 10, wherein the sealed dosage form comprises a dosage strength ofabout 0.5 g, about 0.75 g, about 1.0 g, about 2 g, about 5 g, about 10 gor about 20 g.
 12. The method of claim 9, wherein sterile vancomycinhydrochloride powder is administrable orally or parenterally.
 13. Themethod of claim 1, wherein the spray dried vancomycin hydrochloridepowder has a water content below 3%.
 14. The method of claim 1, furthercomprising: transferring the spray dried vancomycin hydrochloride powderwith EP impurity B level of not more than 1.5% into a gamma irradiatedsingle use bag.